The further we get into mushroom cultivation, the more I realise just how useful and amazing fungi is. I’ve also found that it’s sometimes a little hard to find info that relates to growing edible mushrooms in Australian conditions. Finding local knowledge is crucial!
via Mushroom Cultivation: Good books for Aussies.
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Ganoderma lucidum is a basidiomycete, lamellaless fungus that belongs to the family of polyporaceae. It is also known as Ling Zhi (Chinese), Reishi (Japanese), King of Herbs or God’s Herb. It grows in densely wooded mountains of high humidity and dim lighting. The Polypores (or those of the polyporaceae family), commonly known as brackets or shelf fungi, are prominent mushrooms that grow off the sides of trees.
“Mushroom” is the fruit body of a mycelial tree and has a high-reproductive structure of higher order fungus organism. On the other hand, a mycelium is a network of the threadlike filaments that originate from spores.One of the characteristics of a pure Reishi is that it has an intensely bitter taste.
In modern times, Reishi has been the object of intensive scientific studies to determine its many health benefits from a modern perspective, as it is believed that it has several other major effects in the human body. In fact, continuing researches on the wonders of Ganoderma are not only confined in the Orient – specifically in China, Korea and Japan – but in other Western countries as well.
Its application has been utilized further not only as a supplement but it is also being applied for pharmaceuticals, cosmetics and beverages
Polysaccharides obtained from Ganoderrna lucidum are reported to exhibit
antitumor activity. Recentty, mycelium of this mushroom is being cultivated and the
bioactive polyglycans are also found. We cultured mycelium of G. lucidum cultivated
in Thailand in Potato Dextrose Broth, Yeast-Malt Extract and Glucose-Yeast Extract
liquid media in combination with 20, 40, and 60 gIL glucose. The effects of the media on
mycelial dry weight were studied. Crude extract obtained from mycelium grown in Yeast-
Malt Extract medium were higher than those obtained from other media. It consisted of
significantly higher protein content with low polysaccharide compared to extracts from
. other media. Whereas, crude extract of my~Iium grown in Glucose- Yeast Extract medium
consisted of significantly higher polysaccharide content with low protein. Cytotoxic
activities of hot water extract of mycelia were tested against P-388 leukemia cens in vitro.
Introduction
Ga110derma lucidum, a mushroom widely used in Chinese medicine, is
known for its medicinal values in Asia. The fruiting body and mycelium have been
extracted for active compounds (Jong and Binningham, 1992). Its water extract
polysaccharides are reported to exhibit antitumor activities (Miyazaki and
Nishijirna, 1981; Sone et aI., 1985). Although this mushroom is found’ growing
wildly in Thailand, some strains were also introduced for cultivation.
This paper reports the effects of culture medium on mycelial growth in one
of the cultivated strains and the production of crude extracts with their total
polysaccharides and protein contents. The cytotoxicity effects of these crude extracts
were also studied.
Materials and Method
Mycelium culture
Mycelium of Ganoderma lucidum cultivated in Thailand was kindly
provided by the Research and Development Section, the Royal Chitralada Projects,
Bangkok, Thailand. We cultured mycelium in Potato Dextrose Broth (PDB), Yeastmalt
extract and Glucose-yeast extract liquid medium (Sone et al., 1985) with
varying concentrations of glucose (20, 40 and 60 gIL) under static condition at
28:t2°C. Mycelium dry weights were taken for growth measurement after 28 days in
culture.
Extraction of mycelium
Dry mycelium obtained from each medium was extracted with distilled
water at 1:20 (w/v) at 80°C for 6 h. The extracts were filtered through Whatman
No.1 filter paper and the filtrates were evaporated at 50°C to eliminate
approximately 90% of water. The remaining filtrates were precipitated with absolute
Ethanol at 1:6 (v/v) and incubated at 4°C overnight. Precipitates were collected by
centrifugation at 10,000 rpm, 4°C for 20 min and dried in vacuo.
Determination of total polysaccharides and protein contents
Crude extracts were analysed for total polysaccharide content using Phenolsulfuric
acid method (Dubois et aI., 1956) and for protein content using Lowry’s
method (Dunn, 1995).
Biological assay
MTT (3 -[4,5 -dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide)
colorimeter assay used in this study was modified from Mosmann (1983) and
Jiratchariyakul et af. (1996).
‘BHK (Baby Hamster Kidney) cells were maintained in EMEM medium
(BioWhittaker) supplemented with 10% fetal bovine serum (BioWhittaker) and 100
J.1g/mLpenicillin/streptomycin at 37°C in a humidified atmosphere of 5% CO2. Cells
(2×105 cclls/mL) were inoculated in 50 J.1Laliquot to each well of microtiter plate.
Various sample concentrations (50 J.1L)were added to the cultures at day I after
transplantation. At day 3, the supernatant was removed by carefully inverting,
flicking and blotting the plates. MTT solution (I mg/mL) was added to the cultures
at 50 J.1Lper well. After 4 h incubation, untransforrned MTT was removed and 50
J.1Lisopropanol was added to each well and shake vigorously in a microplate reader.
The optical density measurements were made using a microplate reader at 570 om.
Three replicate wells were used to defermine each ~ata. point. .
P-388 leukemia cells were maintained in RPMI-1640 medium’
(BioWhittaker)supplemented with 10% fetal bovine serum (BioWhittaker) and 100
~mL penicillin/streptomycin at 37°C in a humidified atmosphere of 5% CO2. Cells
(2x I04 cells/mL) were inoculated in 100 J.LLaliquot to each weB of microtiter plate.
Various sample concentrations (100 J.LL)were added to the cultures at day I after
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